CN112174925A - Method for microwave-assisted extraction of grape seed procyanidine - Google Patents
Method for microwave-assisted extraction of grape seed procyanidine Download PDFInfo
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- CN112174925A CN112174925A CN202011350543.8A CN202011350543A CN112174925A CN 112174925 A CN112174925 A CN 112174925A CN 202011350543 A CN202011350543 A CN 202011350543A CN 112174925 A CN112174925 A CN 112174925A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
- C07D311/58—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4
- C07D311/60—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4 with aryl radicals attached in position 2
- C07D311/62—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4 with aryl radicals attached in position 2 with oxygen atoms directly attached in position 3, e.g. anthocyanidins
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D11/00—Solvent extraction
- B01D11/02—Solvent extraction of solids
- B01D11/0288—Applications, solvents
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/10—Selective adsorption, e.g. chromatography characterised by constructional or operational features
- B01D15/20—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to the conditioning of the sorbent material
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/10—Selective adsorption, e.g. chromatography characterised by constructional or operational features
- B01D15/20—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to the conditioning of the sorbent material
- B01D15/203—Equilibration or regeneration
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/42—Selective adsorption, e.g. chromatography characterised by the development mode, e.g. by displacement or by elution
- B01D15/424—Elution mode
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09B—ORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES, e.g. PIGMENTS; MORDANTS; LAKES
- C09B61/00—Dyes of natural origin prepared from natural sources, e.g. vegetable sources
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09B—ORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES, e.g. PIGMENTS; MORDANTS; LAKES
- C09B67/00—Influencing the physical, e.g. the dyeing or printing properties of dyestuffs without chemical reactions, e.g. by treating with solvents grinding or grinding assistants, coating of pigments or dyes; Process features in the making of dyestuff preparations; Dyestuff preparations of a special physical nature, e.g. tablets, films
- C09B67/0096—Purification; Precipitation; Filtration
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- Medicines Containing Plant Substances (AREA)
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Abstract
The invention provides a method for microwave-assisted extraction of grape seed procyanidin, which comprises the following steps: naturally air drying grape seed sample, pulverizing, sieving, adding cellulase and ethanol solution, soaking, extracting with microwave digestion instrument, vacuum filtering, concentrating, freeze drying to obtain crude extract of grape seed procyanidin, adsorbing with AB-8 resin, eluting with ethanol, rotary evaporating for concentrating, freeze drying, and making into powder; the method adopts a microwave and enzyme method double-auxiliary extraction method to extract the procyanidine from the grape seeds, inspects the influence of 3 single factors on the extraction rate of the procyanidine, optimizes the response surface to obtain the optimal extraction condition of the procyanidine, and has wide market prospect.
Description
Technical Field
The invention relates to the technical field of food and processing thereof, in particular to a method for extracting grape seed procyanidine by microwave assistance.
Background
Procyanidins (Procyanidins), also known as condensed tannins, are the second largest natural phenolic substance in nature, next to lignin, present in large quantities in fruits, nuts and seeds, and Procyanidins contained in grape skins and grape seeds are of great interest to research institutions. The oligomeric proanthocyanidin and the polymeric proanthocyanidin are divided into oligomeric proanthocyanidin and polymeric proanthocyanidin according to polymerization degrees, and the powder is generally reddish brown and has little special smell. The procyanidin has good solubility, is soluble in water and organic solvents, and can be rapidly dissolved in most organic solvents.
The existing research shows that the procyanidin has strong antioxidant activity, and the antioxidant capacity of the procyanidin is about 50 times of that of VE and 20 times of that of VC. Procyanidin is the most effective natural antioxidant for scavenging free radicals in human body, and has a large amount of active phenolic hydroxyl groups in procyanidin molecule, which can be combined with double bonds and can be given with hydrogen atoms, so that it can remarkably scavenge free radicals and inhibit lipid peroxidation, and has scavenging ability for free radicals and its stereo structure related. In addition, it can be used for preventing and treating more than 80 diseases caused by free radicals in human body, such as heart disease, arthritis, etc., and has effects of improving microcirculation in vivo, inhibiting bacteria, relieving fatigue, promoting health, and caring skin. Procyanidins also have anti-cancer and anti-tumor activities and cardiovascular patency protecting functions, and are medically used as important components of drugs for preventing and treating cardiovascular diseases.
The existing extraction method of procyanidine mainly comprises the following steps: water-organic solvent extraction, Soxhlet extraction, supercritical CO2 extraction, etc. The organic solvent extraction method has long extraction time and high temperature, and seriously influences the composition of the procyanidine; the Soxhlet extraction method has long time and is difficult to amplify; the supercritical CO2 extraction method has large one-time investment, can not carry out industrialized large-scale production and has high product cost.
Disclosure of Invention
Aiming at the problems, the invention provides a method for extracting grape seed procyanidin by microwave assistance.
The purpose of the invention is realized by adopting the following technical scheme:
a method for extracting grape seed procyanidin by microwave assistance comprises the following steps:
(1) naturally drying the grape seed sample, crushing and sieving with a 80-mesh sieve;
(2) weighing 20g of grape seed powder, adding into a 500mL conical flask, adding 120mL of cellulase with the purity of 80% and 100mL of ethanol solution with the purity of 60%, stirring, and soaking for 30 min;
(3) setting the temperature of a microwave digestion instrument to 60 ℃ in advance, leaching the solution for 10min, taking out and cooling;
(4) carrying out suction filtration, concentration and freeze drying for 20h to obtain an OPC crude extract;
(5) soaking AB-8 resin in industrial ethanol at room temperature for 24h, fully swelling the resin, and washing with distilled water until the effluent liquid has no alcohol smell. The distilled water-washed resin was soaked with a 4% hydrochloric acid solution for 4 hours, and then washed with distilled water until the pH was 7. Then soaking the mixture in 4% sodium hydroxide solution for 4h, and washing the mixture to be neutral by distilled water for later use.
(6) At room temperature, using absolute ethyl alcohol as solvent, standing, sealing and soaking for 8h, then washing with distilled water until no alcohol smell exists, and then soaking with 4% hydrochloric acid and 4% sodium hydroxide solution for standby application when pretreating resin.
(7) Accurately weighing 5g of the treated AB-8 resin, placing the resin in a 250mL conical flask, soaking the resin in 75% ethanol, adding the crude extract of grape seed procyanidin, placing the extract in an oscillator, oscillating for 24 hours at a rotating speed, and taking the supernatant to determine the content of procyanidin.
(8) Filtering the resin adsorbed with procyanidin extract, placing in a conical flask, adding 200mL of 60% ethanol water solution, placing in an oscillator, oscillating, and collecting supernatant to determine procyanidin content.
(9) Concentrating after rotary evaporation, freeze drying, and making into powder.
Preferably, the cellulase is 0.8mg/ml, and the purity of the ethanol is 60%.
Preferably, the microwave temperature is 60 ℃ and the time duration is 10 min.
Preferably, the pretreatment method of the resin comprises the following steps:
soaking AB-8 resin in industrial ethanol at room temperature for 24h, fully swelling the resin, and washing with distilled water until the effluent liquid has no alcohol smell. The distilled water-washed resin was soaked with a 4% hydrochloric acid solution for 4 hours, and then washed with distilled water until the pH was 7. Then soaking the mixture in 4% sodium hydroxide solution for 4h, and washing the mixture to be neutral by distilled water for later use.
Preferably, the resin regeneration method comprises the following steps:
at room temperature, using absolute ethyl alcohol as solvent, standing, sealing and soaking for 8h, then washing with distilled water until no alcohol smell exists, and then soaking with 4% hydrochloric acid and 4% sodium hydroxide solution for standby application when pretreating resin.
Preferably, the required amount of the resin is 5g, the purity of the ethanol is 75%, and the working time of the oscillator is 24 h.
Preferably, the purity of the ethanol for elution is 60%.
The invention has the beneficial effects that:
according to the method, the proanthocyanidins are extracted from the grape seeds by a microwave and enzyme method double-auxiliary extraction method, the influence of 3 single factors on the extraction rate of the proanthocyanidins is investigated, the optimal extraction conditions of the proanthocyanidins are obtained by response surface optimization, the extraction rate is high, the product purity is high, and the market prospect is wide. And also provides scientific basis for efficiently utilizing active ingredients such as procyanidine in wine grape seeds and increasing the added value of industrial production of the wine.
Detailed Description
The invention is further described with reference to the following examples.
The embodiment of the invention relates to a method for extracting grape seed procyanidin by microwave assistance, which comprises the following steps:
(1) naturally drying the grape seed sample, crushing and sieving with a 80-mesh sieve;
(2) weighing 20g of grape seed powder, adding into a 500mL conical flask, adding 120mL of cellulase with the purity of 80% and 100mL of ethanol solution with the purity of 60%, stirring, and soaking for 30 min;
(3) setting the temperature of a microwave digestion instrument to 60 ℃ in advance, leaching the solution for 10min, taking out and cooling;
(4) carrying out suction filtration, concentration and freeze drying for 20h to obtain an OPC crude extract;
(5) soaking AB-8 resin in industrial ethanol at room temperature for 24h, fully swelling the resin, and washing with distilled water until the effluent liquid has no alcohol smell. The distilled water-washed resin was soaked with a 4% hydrochloric acid solution for 4 hours, and then washed with distilled water until the pH was 7. Then soaking the mixture in 4% sodium hydroxide solution for 4h, and washing the mixture to be neutral by distilled water for later use.
(6) At room temperature, using absolute ethyl alcohol as solvent, standing, sealing and soaking for 8h, then washing with distilled water until no alcohol smell exists, and then soaking with 4% hydrochloric acid and 4% sodium hydroxide solution for standby application when pretreating resin.
(7) Accurately weighing 5g of the treated AB-8 resin, placing the resin in a 250mL conical flask, soaking the resin in 75% ethanol, adding the crude extract of grape seed procyanidin, placing the extract in an oscillator, oscillating for 24 hours at a rotating speed, and taking the supernatant to determine the content of procyanidin.
(8) Filtering the resin adsorbed with procyanidin extract, placing in a conical flask, adding 200mL of 60% ethanol water solution, placing in an oscillator, oscillating, and collecting supernatant to determine procyanidin content.
(9) Concentrating after rotary evaporation, freeze drying, and making into powder.
Preferably, the cellulase is 0.8mg/ml, and the purity of the ethanol is 60%.
Preferably, the microwave temperature is 60 ℃ and the time duration is 10 min.
Preferably, the pretreatment method of the resin comprises the following steps:
soaking AB-8 resin in industrial ethanol at room temperature for 24h, fully swelling the resin, and washing with distilled water until the effluent liquid has no alcohol smell. The distilled water-washed resin was soaked with a 4% hydrochloric acid solution for 4 hours, and then washed with distilled water until the pH was 7. Then soaking the mixture in 4% sodium hydroxide solution for 4h, and washing the mixture to be neutral by distilled water for later use.
Preferably, the resin regeneration method comprises the following steps:
at room temperature, using absolute ethyl alcohol as solvent, standing, sealing and soaking for 8h, then washing with distilled water until no alcohol smell exists, and then soaking with 4% hydrochloric acid and 4% sodium hydroxide solution for standby application when pretreating resin.
Preferably, the required amount of the resin is 5g, the purity of the ethanol is 75%, and the working time of the oscillator is 24 h.
Preferably, the purity of the ethanol for elution is 60%.
Example 1
A method for extracting grape seed procyanidin with the assistance of microwave comprises the following steps:
(1) naturally drying the grape seed sample, crushing and sieving with a 80-mesh sieve;
(2) weighing 5 parts of 20g of grape seed powder, respectively adding the grape seed powder into 5 500mL conical flasks, respectively adding 60mL, 90mL, 120mL, 150mL and 180mL of 80% cellulase and 50mL, 75mL, 100mL, 125mL and 150mL of 60% ethanol solution, stirring, and soaking for 30 min;
(3) setting the temperature of a microwave digestion instrument to 60 ℃ in advance, leaching the solutions for 10min respectively, and taking out and cooling;
(4) carrying out suction filtration, concentration and freeze drying for 20h to obtain an OPC crude extract;
(5) soaking AB-8 resin in industrial ethanol at room temperature for 24h, fully swelling the resin, and washing with distilled water until the effluent liquid has no alcohol smell. The distilled water-washed resin was soaked with a 4% hydrochloric acid solution for 4 hours, and then washed with distilled water until the pH was 7. Then soaking the mixture in 4% sodium hydroxide solution for 4h, and washing the mixture to be neutral by distilled water for later use.
(6) At room temperature, using absolute ethyl alcohol as solvent, standing, sealing and soaking for 8h, then washing with distilled water until no alcohol smell exists, and then soaking with 4% hydrochloric acid and 4% sodium hydroxide solution for standby application when pretreating resin.
(7) Accurately weighing 5 parts of treated AB-8 resin 5g, respectively placing in 250mL conical flasks, soaking with 75% ethanol, respectively adding the crude extract of grape seed procyanidin obtained in step (4), placing in an oscillator, oscillating at a rotating speed for 24h, and taking the supernatant to determine the content of procyanidin.
(8) Filtering the resin adsorbed with procyanidin extract, placing in a conical flask, adding 200mL of 60% ethanol water solution, placing in an oscillator, oscillating, and collecting supernatant to determine procyanidin content.
(9) Concentrating after rotary evaporation, freeze drying, and making into powder, which is weighed to be 0.22g, 0.28g, 0.38g, 0.34g, 0.32g respectively.
Preferably, the cellulase is 0.8mg/ml, and the purity of the ethanol is 60%.
Preferably, the microwave temperature is 60 ℃ and the time duration is 10 min.
Preferably, the pretreatment method of the resin comprises the following steps:
soaking AB-8 resin in industrial ethanol at room temperature for 24h, fully swelling the resin, and washing with distilled water until the effluent liquid has no alcohol smell. The distilled water-washed resin was soaked with a 4% hydrochloric acid solution for 4 hours, and then washed with distilled water until the pH was 7. Then soaking the mixture in 4% sodium hydroxide solution for 4h, and washing the mixture to be neutral by distilled water for later use.
Preferably, the resin regeneration method comprises the following steps:
at room temperature, using absolute ethyl alcohol as solvent, standing, sealing and soaking for 8h, then washing with distilled water until no alcohol smell exists, and then soaking with 4% hydrochloric acid and 4% sodium hydroxide solution for standby application when pretreating resin.
Preferably, the required amount of the resin is 5g, the purity of the ethanol is 75%, and the working time of the oscillator is 24 h.
Preferably, the purity of the ethanol for elution is 60%.
Example 2
A method for extracting grape seed procyanidin with the assistance of microwave comprises the following steps:
(1) naturally drying the grape seed sample, crushing and sieving with a 80-mesh sieve;
(2) weighing 5 parts of 20g of grape seed powder, respectively adding the grape seed powder into 5 500mL conical flasks, respectively adding 120mL of cellulase with the purity of 80% and 100mL of ethanol solution with the purity of 50%, 60%, 70%, 80% and 90%, stirring, and soaking for 30 min;
(3) setting the temperature of a microwave digestion instrument to 60 ℃ in advance, leaching the solutions for 10min respectively, and taking out and cooling;
(4) carrying out suction filtration, concentration and freeze drying for 20h to obtain an OPC crude extract;
(5) soaking AB-8 resin in industrial ethanol at room temperature for 24h, fully swelling the resin, and washing with distilled water until the effluent liquid has no alcohol smell. The distilled water-washed resin was soaked with a 4% hydrochloric acid solution for 4 hours, and then washed with distilled water until the pH was 7. Then soaking the mixture in 4% sodium hydroxide solution for 4h, and washing the mixture to be neutral by distilled water for later use.
(6) At room temperature, using absolute ethyl alcohol as solvent, standing, sealing and soaking for 8h, then washing with distilled water until no alcohol smell exists, and then soaking with 4% hydrochloric acid and 4% sodium hydroxide solution for standby application when pretreating resin.
(7) Accurately weighing 5 parts of treated AB-8 resin 5g, respectively placing in 250mL conical flasks, soaking with 75% ethanol, respectively adding the crude extract of grape seed procyanidin obtained in step (4), placing in an oscillator, oscillating at a rotating speed for 24h, and taking the supernatant to determine the content of procyanidin.
(8) Filtering the resin adsorbed with procyanidin extract, placing in a conical flask, adding 200mL of 60% ethanol water solution, placing in an oscillator, oscillating, and collecting supernatant to determine procyanidin content.
(9) Concentrating after rotary evaporation, freeze drying, and making into powder, which is weighed to be 0.37g, 0.48g, 0.42g, 0.34g, 0.30g respectively.
Preferably, the cellulase is 0.8 mg/ml.
Preferably, the microwave temperature is 60 ℃ and the time duration is 10 min.
Preferably, the pretreatment method of the resin comprises the following steps:
soaking AB-8 resin in industrial ethanol at room temperature for 24h, fully swelling the resin, and washing with distilled water until the effluent liquid has no alcohol smell. The distilled water-washed resin was soaked with a 4% hydrochloric acid solution for 4 hours, and then washed with distilled water until the pH was 7. Then soaking the mixture in 4% sodium hydroxide solution for 4h, and washing the mixture to be neutral by distilled water for later use.
Preferably, the resin regeneration method comprises the following steps:
at room temperature, using absolute ethyl alcohol as solvent, standing, sealing and soaking for 8h, then washing with distilled water until no alcohol smell exists, and then soaking with 4% hydrochloric acid and 4% sodium hydroxide solution for standby application when pretreating resin.
Preferably, the required amount of the resin is 5g, the purity of the ethanol is 75%, and the working time of the oscillator is 24 h.
Preferably, the purity of the ethanol for elution is 60%.
Example 3
A method for extracting grape seed procyanidin with the assistance of microwave comprises the following steps:
(1) naturally drying the grape seed sample, crushing and sieving with a 80-mesh sieve;
(2) weighing 5 parts of 20g of grape seed powder, respectively adding the grape seed powder into 5 500mL conical flasks, respectively adding 120mL of cellulase with the purity of 80% and 100mL of ethanol solution with the purity of 60%, stirring, and soaking for 30 min;
(3) setting the temperature of the microwave digestion instrument to 30 ℃, 45 ℃, 60 ℃, 75 ℃ and 90 ℃ in several times, respectively leaching the solution for 10min, taking out and cooling;
(4) carrying out suction filtration, concentration and freeze drying for 20h to obtain an OPC crude extract;
(5) soaking AB-8 resin in industrial ethanol at room temperature for 24h, fully swelling the resin, and washing with distilled water until the effluent liquid has no alcohol smell. The distilled water-washed resin was soaked with a 4% hydrochloric acid solution for 4 hours, and then washed with distilled water until the pH was 7. Then soaking the mixture in 4% sodium hydroxide solution for 4h, and washing the mixture to be neutral by distilled water for later use.
(6) At room temperature, using absolute ethyl alcohol as solvent, standing, sealing and soaking for 8h, then washing with distilled water until no alcohol smell exists, and then soaking with 4% hydrochloric acid and 4% sodium hydroxide solution for standby application when pretreating resin.
(7) Accurately weighing 5 parts of treated AB-8 resin 5g, respectively placing in 250mL conical flasks, soaking with 75% ethanol, respectively adding the crude extract of grape seed procyanidin obtained in step (4), placing in an oscillator, oscillating at a rotating speed for 24h, and taking the supernatant to determine the content of procyanidin.
(8) Filtering the resin adsorbed with procyanidin extract, placing in a conical flask, adding 200mL of 60% ethanol water solution, placing in an oscillator, oscillating, and collecting supernatant to determine procyanidin content.
(9) Concentrating after rotary evaporation, freeze drying, and making into powder, which is weighed to be 0.15g, 0.28g, 0.37g, 0.34g, 0.33g respectively.
Preferably, the cellulase is 0.8 mg/ml.
Preferably, the microwave time is 10 min.
Preferably, the pretreatment method of the resin comprises the following steps:
soaking AB-8 resin in industrial ethanol at room temperature for 24h, fully swelling the resin, and washing with distilled water until the effluent liquid has no alcohol smell. The distilled water-washed resin was soaked with a 4% hydrochloric acid solution for 4 hours, and then washed with distilled water until the pH was 7. Then soaking the mixture in 4% sodium hydroxide solution for 4h, and washing the mixture to be neutral by distilled water for later use.
Preferably, the resin regeneration method comprises the following steps:
at room temperature, using absolute ethyl alcohol as solvent, standing, sealing and soaking for 8h, then washing with distilled water until no alcohol smell exists, and then soaking with 4% hydrochloric acid and 4% sodium hydroxide solution for standby application when pretreating resin.
Preferably, the required amount of the resin is 5g, the purity of the ethanol is 75%, and the working time of the oscillator is 24 h.
Preferably, the purity of the ethanol for elution is 60%.
Finally, it should be noted that the above embodiments are only used for illustrating the technical solutions of the present invention, and not for limiting the protection scope of the present invention, although the present invention is described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions can be made on the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.
Claims (7)
1. A method for extracting grape seed procyanidin under the assistance of microwaves is characterized by comprising the following steps:
(1) naturally drying the grape seed sample, crushing and sieving with a 80-mesh sieve;
(2) weighing 20g of grape seed powder, adding into a 500mL conical flask, adding 120mL of cellulase with the purity of 80% and 100mL of ethanol solution with the purity of 60%, stirring, and soaking for 30 min;
(3) setting the temperature of a microwave digestion instrument to 60 ℃ in advance, leaching the solution for 10min, taking out and cooling;
(4) carrying out suction filtration, concentration and freeze drying for 20h to obtain an OPC crude extract;
(5) soaking AB-8 resin in industrial ethanol at room temperature for 24h, fully swelling the resin, and washing with distilled water until the effluent liquid has no alcohol smell. The distilled water-washed resin was soaked with a 4% hydrochloric acid solution for 4 hours, and then washed with distilled water until the pH was 7. Then soaking the mixture in 4% sodium hydroxide solution for 4h, and washing the mixture to be neutral by distilled water for later use.
(6) At room temperature, using absolute ethyl alcohol as solvent, standing, sealing and soaking for 8h, then washing with distilled water until no alcohol smell exists, and then soaking with 4% hydrochloric acid and 4% sodium hydroxide solution for standby application when pretreating resin.
(7) Accurately weighing 5g of the treated AB-8 resin, placing the resin in a 250mL conical flask, soaking the resin in 75% ethanol, adding the crude extract of grape seed procyanidin, placing the extract in an oscillator, oscillating for 24 hours at a rotating speed, and taking the supernatant to determine the content of procyanidin.
(8) Filtering the resin adsorbed with procyanidin extract, placing in a conical flask, adding 200mL of 60% ethanol water solution, placing in an oscillator, oscillating, and collecting supernatant to determine procyanidin content.
(9) Concentrating after rotary evaporation, freeze drying, and making into powder.
2. The method for microwave-assisted extraction of grape seed procyanidins of claim 1, wherein cellulase is 0.8mg/ml and ethanol purity is 60%.
3. The method for microwave-assisted extraction of grape seed procyanidins of claim 1, wherein the microwave temperature is 60 ℃ and the duration is 10 min.
4. The method for microwave-assisted extraction of grape seed procyanidins of claim 1, wherein the pretreatment method of the resin is as follows:
soaking AB-8 resin in industrial ethanol at room temperature for 24h, fully swelling the resin, and washing with distilled water until the effluent liquid has no alcohol smell. The distilled water-washed resin was soaked with a 4% hydrochloric acid solution for 4 hours, and then washed with distilled water until the pH was 7. Then soaking the mixture in 4% sodium hydroxide solution for 4h, and washing the mixture to be neutral by distilled water for later use.
5. The method for microwave-assisted extraction of grape seed procyanidins of claim 1, wherein the resin regeneration method comprises:
at room temperature, using absolute ethyl alcohol as solvent, standing, sealing and soaking for 8h, then washing with distilled water until no alcohol smell exists, and then soaking with 4% hydrochloric acid and 4% sodium hydroxide solution for standby application when pretreating resin.
6. The method for microwave-assisted extraction of grape seed procyanidin of claim 1, wherein the resin requirement is 5g, the ethanol purity is 75%, and the oscillator working time is 24 h.
7. The method for microwave-assisted extraction of grape seed procyanidins of claim 1, wherein the ethanol used for elution has a purity of 60%.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115715891A (en) * | 2022-11-11 | 2023-02-28 | 湖南绿蔓生物科技股份有限公司 | Continuous method for separating multiple effective components from fructus Siraitiae Grosvenorii |
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2020
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115715891A (en) * | 2022-11-11 | 2023-02-28 | 湖南绿蔓生物科技股份有限公司 | Continuous method for separating multiple effective components from fructus Siraitiae Grosvenorii |
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